胡椒碱对大鼠原代培养肝细胞β-GD活性的影响  

The influence of piperine on β-GD activity of rat hepatocytes in primary culture

作　　者：曾晓会[1] 陈玉兴[1] 黄雪君[1] 杜铁良[1] 赵自明[1] 

机构地区：[1]广东省中医研究所,广州510095

出　　处：《中药药理与临床》2010年第2期32-34,共3页Pharmacology and Clinics of Chinese Materia Medica

基　　金：广东省科技厅火炬计划项目(2005A30101016)

摘　　要：目的:建立原代培养肝细胞模型,观察胡椒碱对肝细胞内β-GD活性的影响,为胡椒碱影响药物吸收和代谢的研究提供一定实验依据。方法:通过门静脉两步灌流法分离原代肝实质细胞后悬浮培养,采用台盼蓝拒染法测定细胞活率,并绘制生长曲线;加入不同浓度的胡椒碱,低温反复冻融细胞,测定上清液中β-GD活性变化。结果:两步灌注分离取得的肝细胞呈类圆形,细胞质透亮,细胞轮廓折光性很强;平均每只大鼠可获得肝细胞1×108个/ml左右,肝细胞存活率为85%;于1、2和3h时,胡椒碱的0.042,0.021,0.0105和0.0052mmol/L均可显著降低大鼠肝细胞里β-GD的OD值(P<0.05～0.01)。结论:门静脉两步灌注法是一种细胞产量高,损伤小,纯度高的分离原代大鼠肝细胞的方法;胡椒碱可抑制大鼠离体肝细胞里β-GD的活性。Purpose：To establish rat hepatocytes model in primary culture,study the influence of piperine on β-GD activity of hepatocytes,and to provide a basis for the impact research of piperine to drug absorption and metabolism experiments. Methods：To apply ＂two-step portal vein perfusion separation＂ into rat model of liver perfusion in situ,in suspension culture and then to proceed the establishment of primary liver cell model; to Assess cell activity then to carry out experiment of the cytotoxic drug; to calculate the maximum non-toxic concentrations and to determine the maximum dose effect for the experiments. And then to take β-GD as the observation indicators to study the reducing difference of β-GD activity. Results：Separation of two-step perfusion of the liver cells were made round,transparent cytoplasm,highly refractive cell contour; The hepatocytes are about 1×108/ml from each rat,its survival was 85%.cytotoxic drugs has showed that when the piperine TC50 is 0.497mg/ml,TC1 is 0.024mg/ml.Compared to the control group,in 1,2 and 3h,piperine could significantly reduce the β-GD activity of hepatocytes. Conclusion：The method of portal vein two-step perfusion is a high injury and the separation of high purity primary rat liver cells; piperine can inhibite β-GD activity of rat hepatocytes in vitro.

关 键 词：胡椒碱 肝细胞 β-GD 

分 类 号：R285.5[医药卫生—中药学]